Simultaneous determination of six bioactive compounds in Evodiae Fructus by high-performance liquid chromatography...

Area of Science:

• Analytical Chemistry
• Pharmacognosy
• Traditional Chinese Medicine

Background:

• Evodiae Fructus (Wuzhuyu) is a traditional Chinese medicine with diverse bioactive components.
• Accurate quantification of these components is crucial for ensuring the quality and efficacy of Wuzhuyu.
• Existing analytical methods may not simultaneously determine multiple key compounds efficiently.

Purpose of the Study:

• To develop and validate a simple, reliable, and simultaneous high-performance liquid chromatography method with diode array detection (HPLC-DAD).
• To quantify six bioactive components: rutaevine, limonin, evodiamine, rutaecarpine, N-formyldihydrorutaecarpine, and dihydroevocarpine in Evodiae Fructus.
• To establish quality control standards for Wuzhuyu based on its key bioactive constituents.

Main Methods:

• HPLC separation using an Agilent Eclipse C18 column at 35°C.
• Gradient elution with a mobile phase consisting of tetrahydrofuran-0.02% phosphoric acid and acetonitrile.
• Diode array detection (DAD) at 220 nm for simultaneous quantification.

Main Results:

• The developed HPLC-DAD method demonstrated excellent linearity (correlation coefficients > 0.999) for all six compounds.
• Average recovery yields ranged from 98.39% to 104.96%, indicating high accuracy.
• The method exhibited excellent repeatability and precision, with relative standard deviation (RSD) < 2.0% for both intra-day and inter-day analyses.
• Successful simultaneous determination of the six target compounds in Evodiae Fructus samples was achieved.

Conclusions:

• The validated HPLC-DAD method is suitable for the simultaneous qualitative and quantitative analysis of multiple bioactive components in Evodiae Fructus.
• This study represents the first simultaneous analysis of indolequinazoline alkaloids, quinolone alkaloid, and limonoids in Wuzhuyu.
• Recommended quality standards for Evodiae Fructus: total evodiamine and rutaecarpine ≥ 0.15%; total rutaevine and limonin ≥ 0.50%.